LOCUS       sequence-74553-e        5410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5410)
  AUTHORS   Rhodius VA, Segall-Shapiro TH, Sharon BD, Ghodasara A, Orlova E, 
            Tabakh H, Burkhardt DH, Clancy K, Peterson TC, Gross CA, Voigt CA
  TITLE     Design of orthogonal genetic switches based on a crosstalk map of 
            sigmas, anti-sigmas, and promoters.
  JOURNAL   Mol Syst Biol. 2013 Oct 29;9:702. doi: 10.1038/msb.2013.58.
  PUBMED    24169405
REFERENCE   2  (bases 1 to 5410)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Syst 
            Biol."; date: "2013-10-29"; volume: "9:702. doi"; pages: " 
            10.1038/msb.2013.58"
COMMENT     Sequence Label: pVRc30_35
FEATURES             Location/Qualifiers
     source          1..5410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(895..912)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1065..1151
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1243..1270
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1289..1380
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1550..2209
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1616..1635)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2351..2806
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2438..2457)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3492..3509)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3626..4171)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3706..3725)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4243..4262
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             5393..5404
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atggataaac gtctgcagca gctgcgtgaa gaatacaaaa atgttcagat tccgaaagaa
       61 ctggatatta ttgtggaaaa agcactgcag caagaaccga aaaaaaaacg tattgttatg
      121 tggccgacca gcgcagcaat tgcagcagca attctgttta ccgcactggt taacattaat
      181 ccggatgcag cacaggcaat gagcaaaatt ccggttattg gcaaaattgt gaaagccatc
      241 acctttattg aaattaaaga agaaaaagat cagagcagca ttgatgtgaa aacaccggca
      301 ctgagcggtc tgagcaataa agaactggaa aatagcatta atgaaaaata tctgaaagaa
      361 agccagcagc tgtataaaga gtttattcag agcaccagca aaaacaaaaa aggccatctg
      421 agcatctata gcgattatga aaccgttacc gatacaccgg atctgctgag cattcgtcgt
      481 aatattgaaa ccacccaggc aagcagctat acccagagcc gttatattac catcgacaaa
      541 aaaaacgata tcctgctgac cctgaaaagc ctgtttaaag atgaacgcta catcaaagtg
      601 atcagccaga acatcaaaga acaaatgaaa cagcagatga aagaagatcc gaataaaatc
      661 tattggctga ccgatgaaga tgccgaaccg tttaaaacca ttctgccgga tcagaccttt
      721 tatatcaccg aagatcataa actggtgatc agcttcgatg aatatgaagt tgcaccgggt
      781 tatatgggtg ttaccgagtt taccattccg accggtgtta ttagcaatct gctggttggt
      841 gaacgttata ttcgttaagg atccctgttt tggcggatga gagaagattt tcagcctgat
      901 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag
      961 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
     1021 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
     1081 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     1141 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
     1201 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg
     1261 atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg
     1321 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagact
     1381 atgagtattc aacatttccg tgtctgctta cataaacagt aatacaaggg gtgtttacta
     1441 gaggttgatc gggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc
     1501 gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     1561 aatcacggga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     1621 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     1681 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     1741 ccgcctgatg aacgctcacc cggagtttcg tatggccatg aaagacggtg agctggtgat
     1801 ctgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcgtc
     1861 cctctggagt gaataccacg acgatttccg gcagtttctc cacatatatt cgcaagatgt
     1921 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     1981 tgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     2041 caacttcttc gcccccgttt tcacgatggg caaatattat acgcaaggcg acaaggtgct
     2101 gatgccgctg gcgatccagg ttcatcatgc cgtttgtgat ggcttccatg tcggccgcat
     2161 gcttaatgaa ttacaacagt actgtgatga gtggcagggc ggggcgtaat aatactagct
     2221 ccggcaaaaa aacgggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat
     2281 tacttcgcgt ttgccacctg acgtctaagc tgtcagacca agtttactca tatatacttt
     2341 agattgattt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2401 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2461 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2521 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     2581 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2641 tttaatagtg gactcttgtt ccaaacttga acaacactca accctatctc gggctattct
     2701 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2761 caaaaattta acgcgaattt taacaaaata ttaacgttta caatttaaaa ggatctaggt
     2821 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2881 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2941 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3001 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3061 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3121 atacctcgct ctgctaatcc tgttaccagt caggcatttg agaagcacac ggtcacactg
     3181 cttccggtag tcaataaacc ggtaaaccag caatagacat aagcggctat ttaacgaccc
     3241 tgccctgaac cgacgaccgg gtcgaatttg ctttcgaatt tctgccattc atccgcttat
     3301 tatcacttat tcaggcgtag caccaggcgt ttaagggcac caataactgc cttaaaaaaa
     3361 gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct
     3421 tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata
     3481 tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa
     3541 atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa
     3601 gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg
     3661 aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt
     3721 ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca
     3781 ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca
     3841 gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta
     3901 tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca
     3961 gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac
     4021 tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt
     4081 ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca
     4141 agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata
     4201 tttgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     4261 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     4321 gatctgctca tgtttgacag cttatcatcg atgcataatg tgcctgtcaa atggacgaag
     4381 cagggattct gcaaacccta tgctactccc tcgagccgtc aattgtctga ttcgttacca
     4441 attattaatt tttaaagtat gggcaatcaa ttgctcctgt taaaattgct ttagaaatac
     4501 tttggcagcg gtttgttgta ttgagtttca tttgcgcatt ggttaaatgg aaagtgacag
     4561 tacgctcact gcaacctaat atttttgaaa tatcccaaga gctttttcct tcgcatgccc
     4621 acgctaaaca ttctttttct cttttggtta aatcgttgtt tgatttatta tttgctatat
     4681 ttatttttcg ataattatca actagagaag gaacaattaa tggtatgttc atacacgcat
     4741 gtaaaaataa actatctata tagttgtctt tttctgaatg tgcaaaacta agcattccga
     4801 agccattgtt agccgtatga atagggaaac taaacccagt gataagacct gatgttttcg
     4861 cttctttaat tacatttgga gattttttat ttacagcatt gttttcaaat atattccaat
     4921 taattggtga atgattggag ttagaataat ctactatagg atcatatttt attaaattag
     4981 cgtcatcata atattgcctc cattttttag ggtaattatc taggattgaa atatcagatt
     5041 taaccataga atgaggataa atgatcgcga gtaaataata ttcacaatgt accattttag
     5101 tcatatcaga taagcattga ttaatatcat tattgcttct acaagcttta attttattaa
     5161 ttattctgta tgtgtcgtcg gcatttatgt ttttcatatt caccaccctg aattgactct
     5221 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gcgcgccgct
     5281 tcgtcaggcc acatagcttt cttgttctga tcggaacgat cgttggctga cctgtaggat
     5341 cgtacaggtt tacgcaagaa aatggtttgt tatagtcgaa taaatactag agaaagagga
     5401 gaaataccat
//
